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ABSTRACT

The work in this thesis concerns the analysis of essential replication genes of the F plasmid of Escherichia coli. The techniques principally employed were the cloning of restriction f ragments, transposition mutagenesis, maxicell analysis

of

proteins, and

DNA

sequencing.

This

work led to the following findings.

1. A 1.15kb region of f5 was shown to contain all the functions required

for

initiation

of F

replication. Insertions of a transposon within this 1.15kb region abolished mini-F replication.

2 . A mini-F plasmid was constructed which constitutes the smallest

F derivative so far reported, This plasmid has an elevated copy number, as a result of deletion of the incC region.

3. At least two proteins specified by the essential F replication region were identified in maxicells. The promoter fragments of two mini-F genes were fused to a B-qalactosidase gene and

demonstrated to have low but significant in vivo activities.

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